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Using a baculovirus display library to identify MHC class I mimotopes.

ABSTRACT: We have developed a baculovirus-based display system for identifying antigen mimotopes for MHC class I-specific T cells. The mouse MHC class I molecule, Dd, was displayed on baculovirus-infected insect cells with a library of 9- and 10-mer peptides tethered via a flexible linker to the N terminus of beta2 microglobulin. As a test case, the library was screened by flow cytometry by using a multimeric fluorescent alphabetaTCR from a mouse T cell specific for Dd plus an unknown self peptide. A mimotope was identified that, when bound to Dd, stimulated the T cell to secret IL-2. The sequence of the mimotope was used to identify a self peptide present in a mouse protein, Spin. The Spin peptide, when complexed with Dd, also activated the T cell. This technique should be generally useful in identifying and manipulating MHC class I peptide mimotopes and epitopes.

SUBMITTER: Wang Y

PROVIDER: S-EPMC548325 | biostudies-literature | 2005 Feb

REPOSITORIES: biostudies-literature

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Using a baculovirus display library to identify MHC class I mimotopes.

Wang Yibing Y Rubtsov Anatoyla A Heiser Ryan R White Janice J Crawford Frances F Marrack Philippa P Kappler John W JW

Proceedings of the National Academy of Sciences of the United States of America 20050207 7

We have developed a baculovirus-based display system for identifying antigen mimotopes for MHC class I-specific T cells. The mouse MHC class I molecule, Dd, was displayed on baculovirus-infected insect cells with a library of 9- and 10-mer peptides tethered via a flexible linker to the N terminus of beta2 microglobulin. As a test case, the library was screened by flow cytometry by using a multimeric fluorescent alphabetaTCR from a mouse T cell specific for Dd plus an unknown self peptide. A mimo ...[more]

PMID: 15699351

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